Selective lighting up of epiberberine alkaloid fluorescence by fluorophore-switching aptamer and stoichiometric targeting of human telomeric DNA G-quadruplex multimer.
Selective lighting up of epiberberine alkaloid fluorescence by fluorophore-switching aptamer and stoichiometric targeting of human telomeric DNA G-quadruplex multimer.
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DOI：
10.1021/ac503730j
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发表时间：
2015-01-06
影响因子：
7.4
通讯作者：
Wang Y
中科院分区：
文献类型：
--
作者：
Zhang L;Liu H;Shao Y;Lin C;Jia H;Chen G;Yang D;Wang Y
适体作为功能元件天然存在于活细胞中，可以将非荧光天然靶标转换为荧光团，在开发高灵敏度和选择性生物传感器和筛选功能剂方面非常有用。这项工作表明，人端粒 G-四链体 (HTG) 可以作为潜在的荧光团转换适体 (FSA) 来靶向天然异喹啉生物碱。我们发现，在此研究的G-四链体和各种结构相似的生物碱中，包括表小檗碱（EPI）、小檗碱（BER）、巴马汀（PAL）、药根碱（JAT）、黄连碱（COP）、瓦尼宁（WOR）、血根碱（SAN）、白屈菜红碱（CHE）和两面针碱（NIT），只有HTG DNA，特别是具有G-四联体四联体 5' 端的 5'-TA-3' 残基 (5'-TAG3(TTAG3)3-3', TA[Q]) 作为最小序列，是选择性点亮 EPI 荧光的最有效的 FSA。与 5' 端侧翼序列相比，四分体的 3' 端侧翼序列对 EPI 识别的贡献明显较小。 EPI 与 TA[Q] (Kd = 37 nM) 的结合亲和力至少比其他生物碱紧密 20 倍。稳态吸收、稳态/时间分辨荧光和 NMR 研究表明，EPI 最有可能以比其他生物碱更特异的方式与核心 [Q] 和 G-四联体四联体之外的 5' 端侧翼序列子结构相互作用。 EPI 与 FSA 的高度选择性和紧密结合以及显着增强的荧光表明选择性 EPI 传感器（检测限为 10 nM）的潜在发展。更重要的是，EPI作为生物碱中最亮的FSA发射体，还可以作为HTG DNA的有效构象探针，并区分DNA G-四链体和RNA对应物。此外，EPI 可以化学计量地结合到长 HTG DNA 多聚体的每个 G 四链体单元，并具有最显着的荧光增强，这是以前报道的探针无法实现的。我们的工作表明 EPI 作为生物成像探针和治疗性 DNA 结合剂的潜在用途。
Aptamers, that exist naturally in living cells as functional elements and can switch nonfluorescent natural targets to fluorophores, are very useful in developing highly sensitive and selective biosensors and screening functional agents. This work demonstrates that human telomeric G-quadruplex (HTG) can serve as a potential fluorophore-switching aptamer (FSA) to target a natural isoquinoline alkaloid. We found that, among the G-quadruplexes studied here and the various structurally similar alkaloids including epiberberine (EPI), berberine (BER), palmatine (PAL), jatrorrhizine (JAT), coptisine (COP), worenine (WOR), sanguinarine (SAN), chelerythrine (CHE), and nitidine (NIT), only the HTG DNA, especially with a 5′-TA-3′ residue at the 5′ end of the G-quadruplex tetrad (5′-TAG3(TTAG3)3-3′, TA[Q]) as the minimal sequence, is the most efficient FSA to selectively light up the EPI fluorescence. Compared to the 5′ end flanking sequences, the 3′ end flanking sequences of the tetrad contribute significantly less to the recognition of EPI. The binding affinity of EPI to TA[Q] (Kd = 37 nM) is at least 20 times tighter than those of the other alkaloids. The steady-state absorption, steady-state/time-resolved fluorescence, and NMR studies demonstrate that EPI most likely interact with the 5′ end flanking sequence substructure beyond the core [Q] and the G-quadruplex tetrad in a much more specific manner than the other alkaloids. The highly selective and tight binding of EPI with the FSA and significantly enhanced fluorescence suggest the potential development of a selective EPI sensor (detection limit of 10 nM). More importantly, EPI, as the brightest FSA emitter among the alkaloids, can also serve as an efficient conformation probe for HTG DNA and discriminate the DNA G-quadruplex from the RNA counterpart. Furthermore, EPI can bind stoichiometrically to each G-quadruplex unit of long HTG DNA multimer with the most significant fluorescence enhancement, which has not been achieved by the previously reported probes. Our work suggests the potential use of EPI as a bioimaging probe and a therapeutic DNA binder.
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